Comparison between two isoforms of glycerol-3-phosphate acyltransferase in microalga Myrmecia incisa: Subcellular localization and role in triacylglycerol synthesis
Comparison between two isoforms of glycerol-3-phosphate acyltransferase in microalga Myrmecia incisa: Subcellular localization and role in triacylglycerol synthesis
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微藻 Myrmecia incisa 中甘油-3-磷酸酰基转移酶的两种亚型的比较：亚细胞定位和在三酰甘油合成中的作用。
DOI：
10.1016/j.algal.2020.102172
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发表时间：
2021-04
期刊：
影响因子：
--
通讯作者：
Sun Zheng
中科院分区：
文献类型：
--
作者：
Sun Liping;Ouyang Longling;Bao Hong;Liu Jianguo;Zhou zhigang;Sun Zheng
微藻是天然存在的丰富的三酰甘油(Tag)来源，微藻衍生的Tag在生物能源、生物材料和其他农业创新方面具有巨大的潜力。甘油-3-磷酸酰基转移酶(GPAT)是催化TAG生物合成的关键酶。此前，我们的实验室已经报道了在含油性藻Myrmecia incisa中首次发现GPAT，在本研究中，从同一藻类中进一步鉴定了一个新的GPAT，命名为MiGPAT2。克隆了MiGPAT2的全长基因，包括1374bp的开放阅读框架、153bp的5‘非编码区和271个编码区的3’非编码区。该蛋白由457个氨基酸组成，具有作为GPAT催化活性中心的HX4D基序。重点比较了两种MiGPATs在亚细胞定位和Tag合成功能方面的差异。分别制备了针对MiGPAT1和MiGPAT2的多克隆和单抗，免疫胶体金标记表明它们分别定位于叶绿体和内质网。GFP融合研究进一步证实了这一点。两种miGPATs均在GPAT缺陷型酵母突变株中异源表达，研究发现，miGPAT2在从头合成Tag中起关键作用，而MiGPAT1更多地参与磷脂的形成，通过间接方式促进Tag的产生。本研究的发现扩大了我们对GPAT成员的功能和进化关系的了解。Incisa，这表明在这种藻类中进行GPAT的基因操作以提高标签产量的可能性。
Microalgae represent a rich and naturally occurring source of triacylglycerol (TAG), and the microalgae-derived TAG has enormous potential in bioenergy, bio-based materials and other agricultural innovations. Glycerol-3-phosphate acyltransferase (GPAT) is a key enzyme that catalyzes the biosynthesis of TAG. Previously our lab has reported the first GPAT in the oleaginous algaMyrmecia incisa, and in the present study, a new GPAT was further identified from the same alga, which was designated as MiGPAT2. A full-length cDNA ofMiGPAT2consisting of a 1374-bp ORF, a 153-bp 5′-UTR and a 271-bp 3′-UTR was cloned. The putative protein composed of 457 amino acids, possessing the HX4D motif that acts as the activity center of GPAT catalysis. Emphasis was put on the comparison between two MiGPATs regarding subcellular localization and TAG synthesis function. A polyclonal and monoclonal antibody were separately prepared for MiGPAT1 and MiGPAT2, and the following immunogold labeling showed they were localized on chloroplast and endoplasmic reticulum (ER), respectively. This was further confirmed by the GFP fusion studies. Both MiGPATs were expressed heterologously in a GPAT-deficient yeast mutant, and it was found that MiGPAT2 played key roles in the de novo TAG biosynthesis, whereas MiGPAT1 was more involved in the phospholipid formation, contributing to TAG production via an indirect way. Findings of the present study expanded our knowledge on functional and evolutionary relationships of GPAT members inM. incisa, suggesting the possibility of genetic manipulation of GPAT in this alga for enhanced TAG production.
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影响因子：
2.9
作者：
Khozin-Goldberg, I;Bigogno, C;Cohen, Z
通讯作者：
Cohen, Z
影响因子：
27.5
作者：
Li, Xiao-Chuan;Zhu, Jun;Yang, Zhong-Nan
通讯作者：
Yang, Zhong-Nan
对微藻 Myrmecia incisa 中 2 型二酰基甘油酰基转移酶的新见解
影响因子：
3.3
作者：
Liu Wei;Sun Zheng;Chen Chun-Xiu;Liu Jian-Guo;Zhou Zhi-Gang
通讯作者：
Zhou Zhi-Gang
DOI：
--
发表时间：
2012
期刊：
--
影响因子：
--
作者：
Zhou Zhi-gang
通讯作者：
Zhou Zhi-gang
影响因子：
3.5
作者：
Hayman, MW;Fawcett, T;Slabas, AR
通讯作者：
Slabas, AR